Objective To determine the interleukin(IL)-17 concentrations in serum and synovi-al fluid from patients with knee osteoarthritis(OA),and evaluate their correlation with disease severity. Methods Serum and synovial fluid were collected from 100 patients with primary knee OA;age-and sex-matched 50 healthy subjects provided serum samples as controls.OA severity and grade were assessed u-sing the Lequesne index and Kellgren and Lawrence(K-L)grading system,respectively.The expression of IL-17 was quantified by enzyme-linked immunosorbent assay.Results Serum IL-17 concentrations were significantly higher in patients than in controls(P <0.05).In the patient group,patients with K-L grade 3 and 4 had higher synovial IL-17 levels than those with K-L grade 2(P <0.05).At the same time,com-pared with K-L grade 3,there was a higher IL-17 level in K-L grade 4 patients(P <0.05).Synovial fluid IL-17 concentration was positively correlated with Lequesne index(r =0.6232,P <0.05).There was no correlation between serum IL-17 concentrations and Lequesne index,however.Conclusion The expres-sion of IL-17 was significantly increased in serum and synovial fluid of knee from patients with knee OA. There was a positive correlation between synovial fluid IL-17 concentrations and OA severity.It suggests that the imbalance of IL-17 expression plays a certain role in the pathogenesis of OA.

[Objective]To explore the therapeutic effects of angelica and sodium ferulate (SF) on experimental osteoarthritis and the related mechanisms in rats. [Method]Animal models of osteoarthritis were established in 40 rats by intra-articular injection of 4% papain. The rats were divided into five groups-model control group,25% and 5% angelica treatment groups, 0.5% and 0.1% SF treatment groups.Another additional 8 rats were used as normal control. Bilateral knees of the animalsin the treatment groups began receiving intra-articular injection of 0.1ml angelica injection or SF of corresponding concentration respectively every 3 days while those of the animals in the control groups received saline. All the animals were sacrificed 6 weeks later and samples of the cartilage, plasma and synovial fluid were taken. The contents of MDA and the activities of SOD in the plasma and synovial fluid were detected.Histological examinations of the cartilage were performed and immunohistochemical analyses for MMP-1, TIMP-1, Bcl-2 and Bax in the cartilage were done.[Result]25% Angelica and 0.5% SF significantly decreased the levels of MDA in both plasma and synovial fluid (P

Medial collateral ligament injury is the most common injury of the knee ligament. Most patients with conservative treatment can achieve preinjury activity level. Nonetheless,it's necessary to eval-uate the severity of the injury and judge the best time and details for surgery as serious injury of the medial collateral ligament will cause chronic knee stability and accelerate its degeneration. This paper summarizes the current status of diagnosis and treatment for medial collateral ligament injury of the knee,aiming to pro-vide guidance for dealing with this lesion.

Objective To explore the effects of different types of passive and active movements of foot and ankle on the venous return in the lower limb in order to provide evidence for prevention of venous stasis and deep- vein thrombosis(DVT)after joint replacement. Methods Peak and average velocities of the femoral vein were detected and measured by using color ultrasound Doppler on 30 limbs in 15 healthy volunteers under the circumstance of resting and different types of passive and active movements of foot and ankle. Results During the passive movements of the flexion and extension, inversion and eversion, and the circumduction of foot and ankle, the flexion and extension as well as the inversion and eversion only slightly increased the velocities of venous blood flow. The circumduction produced a higher increase in venous blood flow than the former two movements, increasing the peak velocity by 31.3％ and average velocity by 33.9％ . In the same 3 types of the active movements of foot and ankle, the active flexion and extension, inversion and eversion resulted in increases in peak velocity of 41.3％ and 32.9％ and in average velocity of 40.1％ and 32.4％ , respectively. The active circumduction of foot and ankle produced the strongest influence on the promotion of venous blood flow with the peak velocity increased by 69.3％ and average velocity by 69.1％ . Conclusion The active circumduction, flexion and extension, inversion and eversion, and the passive circumduction of foot and ankle can be utilized to prevent DVT after joint replacement. The active circumduction of foot and ankle is the optimal exercise to prevent DVT.

BACKGROUND:Basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and bone marrow mesenchymal stem cel s transfected with growth factors can be used as seed cel s in ligament tissue engineering.
OBJECTIVE:To observe biological effect of bone marrow mesenchymal stem cel s transfected with recombinant adenovirus vectors carrying bFGF in three-dimensional co-culture with ligament fibroblasts.
METHODS:Passage 3 bone marrow mesenchymal stem cel s were divided into three groups:control group, Ad-EGFP group and Ad-bFGF group. Under a phase contrast microscope, the changes in cel morphology were observed and the rate of transfection was analyzed by flow cytometry. Proliferation of bone marrow mesenchymal stem cel s and ligament fibroblasts was analyzed by MTS, the expression of bFGF protein in bone marrow mesenchymal stem cel s was determined by ELISA. Scleraxis, col agen type I, col agen type III, decorin and cartilage oligomeric matrix protein levels were detected in BMSCs and ligament fibroblasts using real-time fluorescent quantitative PCR.
RESULTS AND CONCLUSION:Recombinant adenovirus-mediated bFGF gene could transfect bone marrow mesenchymal stem cel s efficiently. After co-culture for 3, 6 days, compared with the control group and Ad-EGFP group, in the Ad-bFGF group, the proliferation ability of bone marrow mesenchymal stem cel s and ligament fibroblasts was enhanced (P<0.01), the expression of bFGF protein in supernatant was obviously higher (P<0.01), the col agen type I, col agen type III, decorin and cartilage oligomeric matrix protein mRNA expression decreased in the ligament fibroblasts (P<0.01), but the mRNA expression of Scleraxis, col agen type I, col agen type III in bone marrow mesenchymal stem cel s increased (P<0.01). The results suggest that the co-culture of Ad-bFGF-transfected bone marrow mesenchymal stem cel s with ligament fibroblasts promotes the proliferation of ligament fibroblasts while decreases the col agen synthesis at the same time, and stimulates the proliferation and differentiation of bone marrow mesenchymal stem cel s into ligament fibrolasts.

Objective To detect type Ⅱ collagen synthesis by cultured human chondrocytes under the influence of different intermittent fluid shearing forces.Methods Second passage human monolayer chondrocytes were divided into low-speed (20 rpm/min), mid-speed (40 rpm/min) and high-speed (60 rpm/min) groups according to the rotation speed of the rocking bed. The expression of type Ⅱ collagen was analyzed using immunohistochemistry and RT-PCRs.Results The absorbance of the mid-speed and high-speed groups was significantly greater than that of the low-speed group and a control group. The average optical density of type Ⅱ collagen by RT-PCR was significantly higher in all three sheared groups than in the control group.Conclusions Intermittent shearing can enhance cellular proliferation and the metabolism of human chondrocytes in vitro and promote the expression of type Ⅱ collagen.

Objective To investigate the effect of recombinant adenovirus-mediated basic fibroblast growth factor (bFGF),interleukin-1 receptor antagonist protein (IL-Ra) and insulin-like growth factor(IGF)-1 gene transfection on human osteoarthritis chondrocytes.Methods Monolayer cultures of human osteoarthritis chondrocytes were transfected with recombinant adenovirus carrying genes encoding the following cytokines: human bFGF,IL-1Ra and IGF-1.Six days later,levels of gene expression and glycosaminoglycan (GAG) in culture supernatant were detected.The proliferation and apoptosis of chondrocytes were analyzed by methyl thiazolyl tetrazolium (MTT) and flow cytometry respectively.Matrix biosynthesis was observed by toluidine blue staining and immunohistochemistry examination of type Ⅱ collagen.The expression of type Ⅱ collagen,MMP-3 and TIMP-1 were analyzed by immunoblotting.Comparisons between groups were performed with one-way ANOVA analysis.Results The expression of all transgenes was high following adenoviral transfection compared with the OA control group (P<0.05).The delivery of bFGF alone promoted the cell proliferation and resulted in a significant enhanced biosynthesis of type Ⅱ collagen and proteoglycans of chondrocytes (P<0.05).Compared with bFGF transfection alone,as two or three of the genes were transfected in different combinations,apoptosis rate of chondrocytes was decreased [(26.1±1.6)%,(19.4±1.0)%.(18.4±1.1)%,(13.9±1.8)%.respectively P<0.05].There was marked enhancement of matrix synthesis and expression of TIMP-1.At the same time,the expression of MMP-3 was inhibited.Conciuslon The bFGF gene transfection mediated with adenoviral vectors can greatly promote cell proliferation and increase matrix synthesis in vitro. Compared to the expression of bFGF alone, concomitant gene transfection of bFGF, IL-IRa and IGF-1 in different combinations plays a complementary role, further enhances matrix synthesis and inhibits matrix degradation.

In the present research, the effects of sintered bone modified with surface mineralization/P24 peptide composite biomaterials on the adhesion, proliferation and osteodifferentiation of MC3T3-E1 cells were investigated. The experiments were divided into three groups due to biomaterials used: Group A (composite materials of sintered bone modified with surface mineralization and P24, a peptide of bone morphogenetic protein-2); Group B (sintered bone modified with surface mineralization) and Group C (sintered bone only). The three groups were observed by scanning electron microscopy (SEM) before the experiments, respectively. Then MC3T3-E1 cells were cultured on the surfaces of the three kinds of material, respectively. The cell adhesion rate was assessed by precipitation method. The proliferative ability of MC3T3-E1 cells were measured with MTT assay. And the ALP staining and measurement of alkaline phosphatase (ALP) activity were performed to assess the differentiation of cells into osteoblasts. The SEM results showed that the materials in the three groups retained the natural pore structure and the pore sizes were in the range between 200-850 μm. The adhesive ratio measurements and MTT assay suggested that adhesion and proliferation of MC3T3-E1 cells in Group A were much higher than those in Group B and Group C (P < 0.05). The ALP staining and ALP activity of MC3T3-E1 cells in Group A were significantly higher than those in Group B and Group C (P < 0.05). The sintered bone modified with surface mineralization/P24 composite material was confirmed to improve the adhesion rate and proliferation and osteodifferentiation of MC3T3-E1 cells, and maintained their morphology.
3T3 Cells ; Animals ; Biocompatible Materials ; Bone Morphogenetic Protein 2 ; Bone and Bones ; Cell Adhesion ; Cell Differentiation ; Mice ; Microscopy, Electron, Scanning ; Osteoblasts ; cytology ; Peptides ; Staining and Labeling

AIM: To investigate the effects and mechanisms of intraarticular injection of doxycycline on experimental osreoarthritis in rabbits. METHODS: An animal model of osteoarthritis in knee of rabbits was established by intraarticular injection of papain. 2 mg or 4 mg of doxycycline was injected (intraarticular) once a day in four weeks. The degeneration of articular cartilage, Mankin s marks of the cartilage tissue, the expression of MMP-13, the release of nitricoxide (NO) and the activity of total nitricoxide synthase (NOS) and inducible nitricoxide synthase (iNOS) of the joint fluid were tested subsequently . RESULTS: In the osteoarthritis model group, the Mankin s marks and the expression of MMP-13 observably increased, and spectrophotometric analysis showed the high concentration of the release of NO and high activity of NOS and iNOS in the joint fluid (P

AIM: To investigate the effects of dehydroepiandrosterone (DHEA) on experimental osteoarthritis in rats. METHODS: Forty rats were randomly divided into four groups. Group A is normal control group. Osteoarthritic models of rats were established by intraarticular injections of papain into the right knee joints of groups B, C and D. Then the right knee joints of rats in groups C and D, respectively, received 150 ?l intraarticular injections of DHEA at a concentration of 50 ?mol?L~(-1) and 100 ?mol?L~(-1), and the right knee joints of rats in groups A and B both received 150 ?l physiological saline, twice weekly for five weeks. Six weeks later, all rats were sacrificed, and the articular cartilage was assessed by gross morphologic, histologic, biochemical and immunohistochemical methods. RESULTS: The cartilage damage in groups C and D was much less than that in group B through observation under a surgical microscope. The Mankin's score, nitric oxide (NO) in the douche of articular cavity, malondialdehyde (MDA) in synovium, the expression of matrix metalloproteinase-1 and 9 in articular cartilage in groups C and D decreased in comparison with group B, and the foregoing indexes in group D decreased significantly compared with group C. However, the activities of superoxide dismutase (SOD) in the douche of articular cavity and blood serum in groups C and D increased in comparison with group B, and the foregoing indexes in group D increased significantly compared with group C. CONCLUSIONS:DHEA shows a cartilage-protecting effect which is in a dosage-dependent manner. The mechanism probably is to inhibit the expression of matrix metalloproteinases and to decrease the release of (NO and enhance the antioxidation.